1-(2-n-propyl-4-amino-5-pyrimidyl-methyl)-2(or 4)-methyl pyridinium salts



United States Patent fifice 3,026,277 Patented Feb. 6, 1962 3,020,277 1-{2-n-PROPYL-4-AMINO-S-PYRMiDYLMETHYD- 2(0R 4)--METHYL PYRIDINIUM SALTS Edward F. Rogers, Middletown, and Lewis Sarett, Princeton, NJ, assignors to Merck 8: Co., line, Rah

way, N.J., a corporation of L cw Jersey No Drawing. Filed July 28, 1959, Ser. No. 829,979 .4 Claims. (Cl. mill-456.4)

This invention relates generally to new chemical coutpounds. More specifically, it is concerned with chemical compounds which are useful in the prevention and the cure of. coccidiosis. Still more particularly, it is concerned with a series of new pyridinium quaternary salts which are efiective in controlling coccidiosis when fed in small amounts to poultry. Still more specifically, it relates to 1 (2 loweralkyl 4 amino pyrimidylmethyl)-lowera.lkyl-pyridinium quaternary salts where the lower alkyl radical in the pyrimidine ring contains at least two carbons, and with methods for preparing such compounds. It is further concerned with novel compositions containing such substances.

This application is a continuation-in-part of the cm pending application of Edward F. Rogers and Lewis H. Sarett, Serial No. 805,688, filed April 13, 1959, and now abandoned.

Coccidiosis is a common and widespread poultry disease caused by' several species of protozoan parasites of the genus Eimeria, such as E. tenella, E. necatrix, E.

acervulirm, E. maxima, E. hagani andE. brunelti. E. tenella is the causative agent of a severe and often fatal infection of the ceca of chickens which is manifested by extensive hemorrhage, accumulation of blood in the ceca, and the passage of blood in the droppings. E. necatrix as well as certain other species attack the small intestine of the chick causing what is known as intestinal coccidiosis. Related species of coccidia such as E. melagridis and E. adenoides are causative organisms of coccidioss in turkeys. When left untreated, the severe forms of coccidiosis lead to poor weight gain, reduced feed efficiency and high mortality in fowl. The elimination or control of coccidiosis is. therefore, of paramount importance in the poultry raising industry.

It has now been found that certain pyridiniurn quaternary compounds are very highly active against the pro tozoa responsible for coccidiosis. An object of the present invention is to provide such novel compounds. Another object is to provide a synthesis of such substances. A further object is the provision of animal feeds and feed supplements and of water soluble compositions containing these pyridinium quaternary salts. Other objects will be apparent from the following discussion of our invention.

According to this invention it has been found that l (2 lon-eralkyl 4 amino 5 pyrimidylmethyl) loweralkyl-pyridinium quaternary salts, wherein the pyridine ring is substituted with from one to three lower alkyl groups and the lower alkyl radical at the 2-position of the pyrimidine ring contains at least two carbon atoms, are very effective in preventing and treating coccidiosis. The preferred compounds of our invention are quaternary salts of the formula ESQ-cures in) wherein R is a lower alkyl radical containing at least two carbon atoms, R is a lower alkyl radical, n has a value of l, 2 or 3, and X is an anion; b and c are posttive numbers having values such that the positive charge of 1) moles of cation is neutralized by c moles of anion X. Thus when X is a monovaient anion such as a halide, b is l and c is 2. When snore than one lower alkyl group is present in the pyridine ring. i.e. when n is 2 or 3, such lower alkyl groups may be the same or different. As described more fully below, these compounds are prepared by reaction of a 2-loweralkyl-4-amino-5- substituted methyl pyrimidine with an appropriately alkylated pyridine. As will be apparent from the above structural formula, the compounds described herein may be considered as substituted pyridines. The pyridine ring is substituted at the l-position by a 2-loweralkyl-4-amino- 5-pyrirnidylmethyl radical; it is further substituted with one or more lower alkyl groups, such as methyl, Cthylv propyl, isopropyl, butyl and amyl groups, such substituents being in at least one of the 2, 3 and/or 4 positions of the ring. The pyrimidine moiety also contains a lower alkyl group at the 2-position of the pyrimidine ring. which group is one containing at least two carbon atoms, e.g. ethyl, propyl, isopropyl, isobutyl and the like. The lower alkyl groups present in the pyrimidine and pyridine portions of these salts need not, of course, be the same in any particular compound.

Although the compounds of the invention are active generally against coccidiosis, certain of the quaternary salts are particularly advantageous in that they possess a broader anticoccidial spectrum, i.e. are active at low levels against more of the most troublesome strains of coccidia, than others, and in that they exhibit minimal undesirable side eifects. The preferred compounds are the 1- (2 ethyl or 2 propyl 4 amino 5 py rimidylmethyl)-loweralkyl pyridinium quaternary salts having a lower alkyl radical at either the 2 or 4 position of the pyridine ring. Included among these are the l-(2- ethyl 4 amino 5 pyrimidylmethyl) 2 methyl pyridinium salts, 1 (2 ethyl 4 amino -5 pyrimidyl- 7 methyl)-4-methyl pyridinium salts, l-(2-propyl-4-amino- S-pyrimidylmethyl)-2-methyl pyridinlum salts, and l-(2 propylA-amino-S-pyrimidylmethyl)-4-mcthyl pyridinium salts, which substances exhibit a very favorable spectrum of anticoccidial activity and therapeutic index. 1-(2- butyl 4 amino 5 pyrimidylmethyl) 2 methylpyridinium salts are likewise preferred since they are particularly effective against E. acervulina.

The anion of the quaternary compound may be an inorganic anion such as chloride, bromide, iodide, nitrate, sulfate, phosphate and the like, or the anion of an organic acid such as citric, tartaric, acetic, picric, stearic, succinic, benzoic, phthalic, phenoxyacetic, embonic, abietic. Z-naphthalene sulfonic or ethylencdiamine tctracctic acids. It may also be the anion of a polymer such as a polyphosphate or polystyrenesultonate ion. The nature of the anion is not critical and any anion may be employed as long at it is not unduly toxic for poultry when the compounds of the invention are employed as coccidiostats. It will be realized by those skilled in this art that an acid addition salt of the primary amino group present in these compounds will also be formed concurrently with the quaternary salt.

The compounds of this invention are synthesized by reacting together an appropriately substituted pyrimidine and a lower alkylated pyridine. As the pyrimidine reactant we may employ an ester of a S-hydroxymethyl pyrimidine and a strong inorganic acid such as a hydrohalic acid. According to one process, a 2-loweralkyl-4-amino-S-halomethyl pyrimidine dihydrohalide, in which the halogen is bromine or chlorine and the lowcralkyl radical contains at least two carbon atoms, is reacted directly with the alkylated pyridine. An excess of the liquid alkylated pyridine or, alternatively, organic solvents inert under the reaction conditions such as lower alltanols, acetonitrile or IIIHLZHX quires el "Qf I RIQCHHQ L l where R is lower alkyl having at least two carbons and R is a lower alkyl radical, n is l3 inclusive and X is a halogen such as chlorine or bromine.

Although the S-halomethyl pyrimidines are generally most conveniently employed for reaction with the alkyl pyridine, the quaternization may also be brought about with other esters of the 2-loweralkyl-4-amino5-hydroxymethyl pyrimidine such as the nitrate ester. Other suitable esters are those of organic sulfinic and sulfonic acids such as the methylsulfinate, p-toluenesulfonate and benzenesulfonate esters.

The quaternization may be conducted so that the particular salt desired is obtained directly. Alternatively, the

quaternary salt recovered from the synthetic reaction medium may be conveniently metathesized to another salt by techniques known in the art.

In using the compounds of this invention in the treatment and prevention of coccidiosis, they are conveniently fed to poultry as'a component of the feed of the animals although they may also be given dissolved or suspended in the drinking water. According to one aspect of the invention, novel compositions are provided in which compounds of Formula 1 above are present as an active anticoccidial ingredient. Such compositions comprise the quaternary salts intimately dispersed in or admixed with an inert carrier or diluent. By 'an inert carrier is meant one that is nonreactive with respect to the quaternary and that may be administered with safety to the animals. The carrier or diluent is preferably one that is or may be an ingredient of the animal feed.

' The compositions which are a preferred feature of this aspect of the invention are the so-called feed suppler tents in which the active ingredient is present in relatively large amounts and which are suitable for addition to the poultry feed either directly or after an intermediate dilution or blending step. Examples of carriers or diluents suitable for such compositions are solid orally ingestable carriers such as distillers dried grains, corn meal, citrus meal, fermentation residues, ground oyster shells, Attapulgus clay, wheat shorts, molasses solubles, corn cob meal, edible vegetable substances, toasted dehulled soya flour, soybean mill feed, antibiotic mycelia, soya grits, crushed limestone and the like. The quaternary salts are intimately dispersed or admixed throughout the solid inert carrier by methods such as grinding, stirring, milling or tumbling. By selecting proper'diluents and by altering the ratio of carrier to active ingredient, compositions of any desired concentration may be prepared. Formulations containing from about 1% to about 40% by weight, and preferably from about 225% by weight of active ingredient are particularly suitable for addition to poultry feeds, and compositions containing from about 5-15 by weight of coccidiostat are very satisfactory. The active compound is normally dispersed or mixed uniformly in the diluent but in some instances may be sorbed on the carrier. The optimal concentration of coccidiostat in these feed supplements will depend to some extent on the particular compound employed. Since it is convenient for the feed manufacturer to use about one pound of feed supplement for each ton of finished feed, the preferred concentration of any one of our coccidiostats in a feed supplement is partly a function of the level of active ingredient desired in he fin shed feed.

Examples of typical feed supplements containing a l- (2-loweralkyl-4-amino-5-pyrimidylmethyl)-loweralkylated pyridinium quaternary salt (where the lower alkyl radical in the pyrimidine ring has at least two carbon atoms) dispersed in a solid inert carrier are:

Lbs.

A. 1-(2-ethyl-4-amino-5-pyrimidylmethyl)-2-methyl pyridinium chloride hydrochloride Wheat standard m ddlings 94,0

B. 1-(2-ethyl-4-arnino-5-pyrimidylmethyl)-4 ethyl pyridinium bromide hydrobromide 10. Corn distillers dried grains 90.0

C. 1-(2-ethyl-4-amino-5-pyrimidylmethyl)-2-methyl- S-ethyl pyridinium chloride hydrochloride 15.0 wheat standard middlings 51 D. l-(2-ethyl-4-amino 5 pyrimidylmethyl)-2,3-dimethylpyridinium bromide hydrobromide 12.0 Molasses sol 88.0 E. l-(2-n-propyl-4-amino 5 pyrimidylmethyl)-2- methyl pyridinium chloride hydrochloride 20.0 Corn germ mea 30.0 Corn distillers gr 50.0 F. 1-(2-ethyl-4-amino-5-pyrimidiylmethyl)-2 methyl pyridinium embonate 4O 0 Soya gr 60.0 G. l-(2-ethyl-4 amino 5 pyrimidylmethyl)-4 npropylpyridinium sulfate Fermentation residues 50.0 Wheat shor 45.0

H. 1-(2-n-propyl 4 amino-5-pyrimidylmethyD-4- methyl pyridinium bromide hydrobromide 12.0 Wheat standard dlings 3&0

These and similar feed supplements are prepared by uniformly mixing the quaternary salt with the carrier or carriers.

The feed supplements of the type illustrated hereinabove are usually further diluted with materials such as corn meal or soybean meal before being incorporated in the animalfecd. In this intermediate processing step the level of coccidiostat in the carrier is brought down to from about 0.1% to about 1.0% by weight. This dilution serves to facilitate uniform distribution of the substance in the finished feed. The finished feed is one that contains a source of fat, protein, carbohydrate, minerals, vitamins and other nutritional factors.

The amount of pyridinium quaternary salt required for optimum control of coccidiosis in poultry will, of course, vary somewhat with the specific compound or compounds empolyed. The compounds of Formula I above are efl'ective when administered at levels of less than about 0.05% by weight of the feed. With the preferred compounds of the invention, i.e. the l-(2-cthyl- 4-amino-5-pyrimidylmethyl)-2(or 4)-loweralkyl pyridinium and 1-(2-propyl-4-arnino-S-pyrimidylmethyl)-2(or 4)-loweralltyl pyridinium salts, good prophylactic results are obtained when from about 0.0005% to about 0.05% by weight of the total feed consumed is administered; for most satisfactory results from the standpoint of both efficacy and incidence of undesirable side effects it is preferred that the poultry feed contain between about 0.0025% and 0.025% by weight of pyridinium salt. When the pyridinium salts are to be employed as therapeutic agents, the higher levels may be used for relaoutbreak of coccidiosis. It is desirable to employ the lowest levels that afford fully adequate control of coccidiosis in order to eliminate as far as possibleany risk of side eti'ects that might appear on prolonged feeding of the compounds.

Many of these pyridinium quaternary salts are desirably or advantageouslyadministered to poultry by way ..of the drinking water of the birds. This method of treatment is often employed in the therapestic use of our compounds since poultry with coccidiosis are apt to consume less solid feed than normal birds. The watersoluble. quaternary salts may be added directly tothe drinking fwater; Alternatively; water-soluble powders may be prepared, in which the coccidiostat is intimately admixed with 'a suitable carrier. such as dcrtrose or sucrose, and these powdersaddedto the drinltiig water of poultry as necessary. Such water-soluble ipowders may contain: any desired'concentration of coccidiostat,

and preparations containing from 1-25% by weight of active compound are suitablev Thejfoll'owing'examples are given for purposes of illustration andnotby way of limitation:

A- A slurry of 100 grams of 2 ethyl-4amino-5-brornomethylpyrimidine dihydrobromide in500 ml. of acetoni- 'trile was stirred at room temperature and 100 ml. of 2- methyl pyridine (u-picoline) was added as rapidly as possible. I =temperature rose from -25 C. to 35' C.

' amino-5 -pyrimidylmethyl)-2-rnethyl pyridinium bromide hydrobromidestarted to precipitate in about. a minute.

- The reaction mixture, after-stirring overnight at room The solution became almost clear, and the l-(2-ethyl-'4- temperature under. nitrogen, was filtered, the solid washed with 100 ml. of acetonitrile and two 100ml. portions 4 T of ether, and dried undervacuum at 40 C. The 1-(2- ethyl-4 amino-5-pyrimidylmethyl)-2-methyl pyridininm bromide hydrobromide thus obtained weighed?) grams,

meiun poimtzss-zrot C.'(de c.).

two litersof 2.5 N hydrochlorio acid was put in a column' I and backwashed with distilled water to remove fine solids. The resin' was then washedwith 'distilled'water until the eluate wasless than 0.05 N-in' hydrochloric acid. The

' pyridinium bromide obtained in. Part A above was dis: solved ingtwo-volnnies of water-and the solution put on the resin column. The flow rate was adjusted to ml. per minute, and after a displacement cut of about 300 ml., three product cuts of 200 ml. each were taken. The presence of productin the eluate was indicated by color,

' striations,-and halide titer. The three product cuts were combined when potentiometric titration with silver nitrate indicated the absence of bromide ion, and concentrated .imder' vacuum to a slurry of about 100 ml. The slurry dinium bromide hydrobromidc thus obtained melt at about 270 C. (dec.).

EXAMPLE 3 One gram of 2-propyl-4-amino-5-pyrimidylmethyl bromide dihydrobromide was dissolved or suspended in 7 ml. of dry dimethylformamide, and to this solution was added 5 ml. of 2-methyl pyridine. A clear solution formed from which crystals of l-(2-propyl-4-amino-5- pyrimidyl-methyD-Z-methyl pyr'idinium bromide hydrobromide soon precipitated. These were collected by filtration and dried, melting point 242-3 "Y C. (dec.). The material was purified by recrystallization'from a methanol-ethanol mixture.

EXAMPLE 4.

To a suspension of 7.6 grams of Z-ethyl-Lamino-S- pyrimidylmethyl bromide dihydrobromide i; 50 ml. of V acetonitrile was added 9 ml. of 2mcthyl5-ethyl pyridine. This mixture became warm and a clear solution resulted. After a short time 1-(2-ethyl4-amino-5-pyrimidylmethyl)-2-methyl-5-ethyl pyridinium bromide hydrobromide began to crystallize. Crystallization 'was continued overnight and the white crystals then' collected by filtration, melting point 252' C; (dec.) The crystals were dissolved in 50 ml. of methanol and the solution then diluted with 150 ml. of acetone. The crystals of product thus obtained melted at 261 C. (dec.). 7

EXAMPLE 5 1.9 grams of p-toluene sulfonyl chloride was added gradually with shaking to a cooled (0-5 C.) solution of 1.67 grams of 2-n-propyl-4-amino-S-hydroxy-rnethyl py-- rimidine in 10 m1- of Z-rnethylpyridine. The reaction mixture, after standing threevhours in an ice bath, and 15 hours at room'temperature, was evaporated to dryness in vacuo. .The residue was dissolved in 20 ml. of water,

. acidified with hydrochloric acid and poured over a column was. stirred at room temperature and diluted with two litersof acetone. After aging for two hours in an ice pyridinium 'chloride' hydrochloride was filtered off, washed with two 100ml. portions of acetone and dried under vacuum at 40 C. Yield 73 grams, 95% of theory. U.V. (0.1 M HCl) A 2460, E% 448; A 26%,

' 13% 4,33. Melting point 266-267 c. (deci).

V 7 EXAMPLE 2 v To 5 ml. of 2-methylpyridine (e-picoline) was added a solution of 2 grams of Z-ethylA-amino-S-pyrirnidyl- 3 methyl bromide-dihydrobromide in .15 m1. of methanol.

A clear solution-formed which became slightly warm.

.In a short time crystals of 1-(2-ethyl-4-amino-5-pyrimi dylmethyl)-2-methylpyridinium bromide hydrobromide began to appear. .The mixture was allowed to stand for :about lf hours and then a mixture of ether-petroleum etheradded to complete the crystallization. The crystal-v line material was then collected by filtration, and re-- crystallized iron; 18-20 ml. of methanol. The crystals of 1-(2-ethyl-4-amino-5-pyrimidylmethyl)- 2 -methylpyriof Amberlite IRA- 400 ion exchange resin on the chloride cycle. The eluate was evaporated to dryness to give a slowlydiluted with about 8 ml. of acetone. The salt crys-' tallizcd and on drying melted at 279-281 (dec.).

' EXAMPLE 7 5.1 grams of Z-cthyl-4-amino-5-chloromethyl pyrimidine hydrochloride and 25 ml. of Z-mcthyl pyridine were mixed together in a 50 ml. flask and the mixture warmed on a steam bath. with stirring for two hours. The mixture was then allowed to stand at room temperature for about 15 hours. The solid l-(2-ethyl-4-amino-5-pyrimidylmethyl)-2-methyl pyridinium chloride hydrochloride thus obtained was recovered by filtration, washed with acetone and air dried. 7.2 grams of quaternary salt were obtained, melting point 260-261? C. (dec.).

EXAMPLE 8 The pyridinium quaternary salts set forth in Table l below were synthesized utilizing processes described in preceding examples. In most experiments a 2-loweralkyl- 4-amino-5-halomethylpyrimidine was employed as the pyrimidine reactant, although in'soine cases an ester of a S-hydroxymethyl pyrimidinewas utilized.

days following the date of infection. At the end of this time the infected birds still living were sacrificed. The oocyst count was determined by a microscopic examination of the cecal homogenates.

The results employing the indicated amounts of quaternary salt, and expressed as mean values, are set forth in Table II.

Table I Nmmx r-r1=n+ R R R R r.

CH -CH +N R 2X- N X N R J r. I! I I a 3 R r-.' R R N R i Quaternary Meltli': Cpd. R R R 8* R R X Prepared Point Percent No. By Method (dec.) Feed 1 of Ex. No.

2 266-270 0. 001 6 279-281 0. 001 4 228 0.003 6 253 0. 003 01H: 4 261 0. 002 05H; 6 269 0. 002 4 244 0.002 4 2X) 0. 01 4 267 0.006 4 241 0. 003 CH; 4 239 0.003 4 257 0.006 4 252 0. 003 12 281-283 0.003 3 242-243 0. 003 5 246 0.003 -3 244 0.006 CH1 5 236 0. 01 12 mean 0. 003 12 258-260 0.0125 12 244-246 0.006 4 233 0. 0125 Br]...t i1 205 0603345 e 10 "1103553. embnute 10 0. 003 naphthalene 10 0. 004

ulfonote.

B through R are hydrogen unless specified otherwise. b mtlibmdlwlvity of the pyridinium quaternary salts described in the above table: was determined y o I .Stmht run White Leghorn chicks, in groups of three each, were weighed and placed in cages with wire floors. hey were led ad libitum a standard laboratory basal ration in which graded levels tthe yridinium quaternary salt were intimately dispersed or blended. Normal and infected control birds we: 3 fed 1 ration omtaining no quaternary salt On the second day of test, the chicks were each 0 y mocLlated with 50,000 sporniatod oocysts of E'z'mena tendla. Papers under the cages were examined on the sixth, seventh and eighth days oi may for the presence or absence of bloody droppings. A score of 0 was given it no bloody spots were observed; scores of 1. 2 or 4 were assigned for 1-3, 4-6, and 6 bloody spots, respectively. On the eighth day of my, the surviving birds were weighed, sacrificed and examined grossly for cecal ooccidiosis lesions. Normal tees ere scored 0. and new with detectable, moderate or maximal lesions were scored 1, 2 and 4, respectively. bm a bird died and ceml cocudlosis lesions were present, a score of was recorded. If the total 0! the two mes was '-5, tin compound under test was rated active"; ii the total score was 6 or more the compound was rated muss-e" at the mnoentration tested.

The compounds des'u-ibed in Table I, when tested by this method were found to be highly active (total score 6 0-5) at the dose level indicated.

EXAMPLE 9 1 Table '50 The coecidiostatio activity of two of the pyridinium Pam; of compounds of the invention was determined by the fol-' Percent Percent o vsts lowing mcthod Compound pound Mor- Weight X10" in In Feed taiity Gain Surviving Groups of ten two-week old White Leghorn chicks were fed a mash diet containing various amounts of the compounds. The 7 compounds were uniformly dispersed in 1 ethyl 4- wny- 0015 -0 90 a the feed. After having been on this ration for 24 hours, fifig fikfifi g'fiafi figl 8% 8 33 2%: each chick was inoculated with 50,000 sporulated drochicride 0.0125 0 100 0. 1 f E "a 1 (2 ethyl 4 amino-5-py- 0. 0015 0 97 0.2 OOCYSlSO tene rlrnidyhnethyl 2-methyl- 0.003 0 97 0. 1 Other groups of ten chicks each were fed a sumlar gawk-$225 32? 'figg 95 mash diet which contained-no coccidiostat. These were mm Controls 4 2; also infected after 24 hours and served as positive or Norm! 00mm" 0 90 infected controls. As positive controls, two to four groups of ten chicks each were employed. other groups of ten chicks each were fed the mash free of coccidiostat and s5 EXAMPLE 10 were not infected with coccidiosis. These served as norm1 I ls. A. Three grams of 1-( Z-ethyl-iammo-S-pynmrdyl- The diets were administered to the chicks for eight methyl)z'methyl Pyndmmm bmmldc hydmbromldg 20 ml. of water was treated with N calcium picrate solution until precipitation appeared to be complete. The solid 1-(2-ethyl-4-amino 5 pyrimidyhnethyl)-2-methyl pyridinium dipicrate thus obtained was recovered by filtration. It was purified by dissolving in 15 ml. of hot dimethyliormamide, filtering said solution, and reprecipitsting the salt by treatment of the filtrate with ml. of

' methanol. On drying the product had a melting point of 173--4 C. (dec.),

B. '1-(2sethyl-4-amino-5-pyrimidylmethyl 2 methyl pyridinium embonate was prepared as follows: Two grams of the pyridinium bromide hydrobromide in 10 ml. of water was added to a hot solution of 4.4 grams of sodium cr'nbonate in-75 ml. of water. The l-(2-ethyl-4-amino-5- "pyrimidylmethyD-Z-methyl pyridiniurn embonate precipi' t'at ed immediately, and was collected by filtration and washed with water. It was recrystallized from 100ml. of methanol to give substantially pure material, melting point 200 C. (dec.). 1

c. 3.01 grams r 1-(2-ethyl-4-amino-5-pyrimidylmethyl)-2-methyl pyridinium chloride hydrochloride was dissolved'in '10 ml. of water. To this solution was added 3.5 grams of naphthalenelj-disulfonieacid. The solution remained clear'for a few seconds and then crystal- I lization of the naphthalene disulfonate quaternarysalt began. The reaction mixture was allowed to stand in ice water for two hours. At the end of which time, the l- (2-ethyl-4-amino s pyrimidylrnethyl)-2-methyl pyridinium-LS-narihthalene disulfonate was recovered by filtration and washed with water. It was obtained as colorless crystals, melting point 266-268" C. (dec.).

D. Fifteen grams of Amberlite Ill- 120 resin was charged to a glass tube to give a resin column of about 18 cm. in height. The resin was converted to the hydrogen form by washing with dilute hydrochloric acid and then with water until the effiuent had a pH of 5-6.

V 7.52 grams er l-(2-ethyl-4-amino-5-pyrimidylinethyl) '2-methyl pyridinium chloride hydrochloride in 325 ml. of

pyrimidylmethyD-S-methyl pyridinium bromide hydrobrornide, melting point 281-283 C. (dec.),

EXAMPLE 13 To a stirred suspension of 20 grams of 2 elhyl-4-amino- Spyrimidylmethyl bromide dihydrobromidein l ml. of acetonitrile was added 25 ml. of 4-methylpyridine. The mixture became very warm and there was nearly a complete solation when a new solid began to crystallize.

After standing for 16 hours, 19 grams of l-(2-ethyl-4- amino-S-pyrimidylmethyl)-4-methyl pyridinium bromide hydrobromide was obtained. 0n recrystallizing irom methanol-acetone it had melting point 252' C. (dec.).

EXAMPLE 14 h To a stirred suspension of 20 grams of 2-ethyl-4-amino water wasp'assed over the column for a period of about two hours. The resin was then washed with 125 ml. of

, water and dried invacuo at C." Byanalysis of the effiuentit was determined that 7.15 grams of the pyridinium salt was absorbed on the resin. There was obtained 19.87 grams of l-(2-ethyl- 4-amino-5-pyrimidyl- EXAMPLE 11 15 grams of 2-amyl-4-amino-5-bromomethyl pyrimidine dihydrobromide in 20 ml. of methanol was mixed with S-pyrimidylmethyl bromide dihydrobrornidein lflll ml. of

acetonitrile was added 30ml. of 4-n-propyl pyridine.

Whennearly all the solid had dissolved a new'crystalline material began to crystallize. After'l6hours the crystalline product, l-(2 ethyl-4-amino-5-pyrimidylmethyl)-4 .propyl pyridinium bromide hydrobromide, was collected.- It was recrystallized from ethanol-acetone,. tnelting point .35 3

246 C. '(dec.).' I v EXAMPLE l6 The 2-loweralkyl-4-amino-5-halomethyl pyrimidines employed in making the quaternary compounds of this in- 15 ml. of m-picoline. 100 ml. of isopropanol was added to the reaction mixture and the whole allowed to stand overnight at room temperature. Ether was then added to complete crystallization and the crystals of 1- (2-amy1- 4-arnino-5-pyrimidylmethyl)-2 methyl pyridinium bromide hydrobromide recovered by filtration. On recrystallization from ethanol-ether, the product melted at 205 C. (dec.).- 1

EXAMPLE 12 To a suspension of 20 grams of 2-n-propyl-4-amino-5- bromomethyl pyrimidine dihydrobromide in 120 ml. of

acetonitrile was added 14 ml. of 4-methylpyridine. The mixture became clear after shaking for a few minutes and in a short time crystals began to form. The reactior. mixture was allowed to stand overnight. The resuiting crystals of 1-(2-n-propyl-4-amino-5-pyrimidylvention are prepared by methods described in the literature or in the following manner:

A. 2-l0weraIkyl-4-amino-5-cydnopyrimidine.-A slurry of 54.7 grams of butyramidine hydrochloride and ml. of absolute ethanol is treated at about 5 .C.*with a solution of 11 grams of sodium in 220 ml. of absolute ethanol. The resulting solution is added with stirring at 10-12 C. over a thirty minute period to 53.7 grams of ethoxymethylenemalononitrile in 54 ml. of absolute ethanol. The resulting mixture is stirred at 0 C. for six hours and then at room temperature-for about 12 hours. The mixture is then filtered, evaporated to dryness in vacuo and the residue treated with water. The alcoholic and aqueous solution precipitates are combined, washed with water n-propyl 4 amino 5 pyrimidylmethyl)-4-n-propyl and dried in air. The product is recrystallized from alcohol to give 2-n-propyl-4-aminoJ-cyanopyrimidine, melting point 158-160 C, l

When the above reaction is carried out with isobutyramidine there is obtained 2-isopropyl-4-amino-5- cyanopyrimidine, melting point ISO-151 C. When amylamidine is employed as starting material the end product is 2-butyl-4-amino-S-cyanopyrimidine, melting point Mil-147 C. When hexylamidine is utilized as starting compound, there is obtained 2-amyl-4-amino-5- cyanopyrimidine, melting point 149-450 C.

B. 2-Ioweralkyl-4-amino-5-aminomethylpyrimidin di-- hydr0chloride.l6.2 grams of 2-n-propyl-4-amino-5- cyanopyrimidine is reduced at about 40 lbs. pressure in 200 ml. of methanol in the presence of 26 grams of ammonia and one tablespoon of Raney nickel. The drop in pressure is about 36.5 lbs. The reaction mixture on completion of the reduction is concentrated to a syrup after filtering off the catalyst. The residue thus obtained is acidified with dilute hydrochloric acid and concentrated to a crystalline mass. On recrystallization from methanolacetone there is obtained 2-n-propyl 4-amino-5- amino-methylpyrimidine dihydrochloride, melting point When the 2-isopropyl, Z-butyl and 2-amyl-4amino-S .cyanopyrimidines obtained as described above are used as starting materials in this reduction, there are obtained respectively 2 isopropyl-4-amino-5-aminon1ethylpyrimidine dihy'drochloridg'mtlting point 257-260 0., 2-butyl- 4-amino-5-an1inomethylpyrimidine dihydrochloride, melting point 221-223. 0., and 2-amyl-4-amin0-S-aminomethylpyrimidine dihydrochloride, melting point 188- 189" C. C. 2 lowernlkyl 4-amino-5-hydroxy methyl pyrimi- 'dine.'lwelve grams of 2-n-propyl4-amino-5-amino methylpyrimidine dihydrochloride in 50 ml. of water is treated at 50-60'C. with a solution of 3.5 grams of sodium nitrite in 30 ml. of water.

The sodium nitrite is added dropwise over a, 45 minute period. The heating is continued for an additional two hours, and the reaction mixture then made alkaline with sodium carbonate and extracted with butanol." The butanol extract is evaporated to dryness and the residue crystallized from acetone to give 2-n-propyl-4-amino-5.-hydroxy methylpyrimidines melting point 115-6 C. V

When the Z-isopropyl, Z-butyl and 2-amyl-4-aminoamino methylpyrimidine dihydrochlorides obtained as in Part B above. are utilized in this reaction in place of the I Z-n-propylcompounds, there are obtained 2-isopropyl-4- amino-S-hydroxy methylpyrimidine, 2-bntyl-4-amino-5- hydroxy methylpyrimidine and 2 amyl-4-amino-5-hydroxy methylpyrimidine.

D. Z-Ibweralkyl-fiamino-S-broma methylpyn'midine. The 2-n-propyl-4-amino-5-hydroxy methylpyrimidine obtained in Part C above is dissolved in 15 m1. of acetic acid saturated with hydrogen bromide, andthe mixture 12 bromo methylpyrimidine dihydrobromide, melting point 191-2 (3., 2-butyl-4-amino-5-bromo methylpyrimidine dihydrobromide, melting point l45l50 C., and 2-amyl-4- amino-S-bromo methylpyrimidine dih drobromide.

EXAMPLE 17 When the quaternary salts of Examples 12 through 15 are treated with hydrochloric acid by the method of Example 6, the corresponding chloride hydrochloride quaternary salts are obtained.

Any departure from the above description which conforms to the present invention is intended to be included within the scope of the claims.

' 4-amino-5-pyrimidylmethyl)-4-methyl What is claimed is:

1. A member of the group consisting of l-(2-n-propyl- 4-amino-5-pyrimidylmethyl)-2-methyl pyridiniurn quaternary salt, wherein the anion of the quaternary salt is a non-toxic anion; and non-toxic acid addition salts thereof.

2. A member of the group consisting of 1-(2-npropylpyridinium quaternary salt, wherein the anion of the quaternary salt is a non-toxic anion; and non-toxic acid addition salts thereof.

3. 1-(2-n-propyl-4-amino-5-pyrimidylmethyl)-2-rnethyl pyridinium chloride hydrochloride.

4. 1-(2-n-propyl-4-amino-5-pyrimidylmethyl) -4-methyl pyridinium chloride hydrochloride.

References Cited in the file of this patent UNITED STATES PATENTS Wilson Feb. 26, 1952 2,823,160 Lux d a] Feb. 11, 1958 2,895,874 Lux et a1; July 21, 1958 OTHER REFERENCES Finkelstem et al.: Jour. Org. Chem, vol. 4, pages 365- (1939).

Dornow et al.: Chem. Berichte, vol. 80, pages 502-5 (1947). r

Fujita et al.: Journ. of Biol. Chem, vol. 196, pages 297-303, pages 313-320 (1952).

Rodd: Chemistry oi Carbon Compounds, vol. IVa, page 518 (1957), 

1. A MEMBER OF THE GROUP CONSISTING OF 1-(2-N-PROPYL4-AMINO-5-PYRIMIDYLMETHYL)-2-METHYL PYRIDINIUM QUATERNARY SALT, WHEREIN THE ANION OF THE QUATERNARY SALT IS A NON-TOXIC ANION; AND NON-TOXIC ACID ADDITION SALTS THEREOF. 